关键词: NDR2;基因，Myc表达;免疫血清;亲和纯化;易感性与特异性

    摘 要:目的 制备高效价的NDR2抗体. 方法 构建pRset-A-ndr2，pGEX-4T-1-ndr2-a和pGEX-4T-1-ndr2-b三种重组表达质粒，并分别在大肠杆菌中诱导表达相应的融合蛋白;用全长GST-NDR2蛋白免疫兔，然后用GST-NDR2-A和GST-NDR2-B片段加强免疫;对包涵体形式表达的6His-NDR2进行初步的纯化;利用固定于硝酸纤维素膜上的NDR2抗原亲和吸附纯化抗血清. 结果 经免疫得到了高效价的兔抗人NDR2多克隆抗血清，亲和纯化提高了NDR2抗体的特异性;免疫组化表明NDR2是一种胞浆蛋白. 结论 用全长NDR2蛋白免疫兔，再用片段加强免疫，可以提高蛋白的抗原性，制备得到高效价的抗体.纯化后的特异性NDR2抗体，为进一步研究NDR2的功能打下了必要的基础.

    Preparation，purification and identification of rab┐bit antiserum against human NDR2protein

    ZHANG Wen-Hong，LIU Xing-Ping，WANG Ji-Cun，HAN Yue-Heng，DENG Yan-Chun，ZHOU Jie，YAO Li-Bo

    Department of Biochemistry and Molecular Biology，Fourth Military Medical University，Xi'an710033，China

    Keywords:n-myc downsream regulator2;genes，myc;gene sensitivity and expression;immune sera;affinity purification;specificity

    Abstract:AIM To obtain an anti-NDR2polyclonal anti-body.METHODS Full length of ndr2cDNA was cloned in-to pRset-A vector and two truncated sequences of ndr2were cloned into pGEX-4T-1vector respectively.The fusion pro-teins were expressed in E.coli and partially purified by inclu-sion body fraction.Immunization in rabbits with the whole NDR2protein was reinforced with two truncated fragments of NDR2twice.The anti-NDR2specific antibody was puri-fied by absorbing antiserum with NDR2antigen immobilized NC filter.RESULTS High titer of antiserum against NDR2was obtained in immunized rabbits and the specificity to NDR2was increased after affinity purification.Immunohis-tochemical test with this antibody showed that NDR2is a cy-toplasmic protein.CONCLUSION Reinforcement with trun-cated NDR2proteins can strengthen the antigenicity of NDR2protein in rabbits.The specific NDR2antibody can be ob-tained by NC filter affinity purification. ......